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通过 CRISPR/Cas9 在 中敲除 UDP-糖基转移酶基因揭示了其在杀虫剂解毒中的作用。

Knocking Out of UDP-Glycosyltransferase Gene via CRISPR/Cas9 in Reveals Its Function in Detoxification of Insecticides.

机构信息

Innovation Center of Pesticide Research, Department of Applied Chemistry, College of Sciences, China Agricultural University, Beijing 100193, China.

National-Regional Joint Engineering Research Center for Soil Pollution Control and Remediation in South China, Guangdong Key Laboratory of Integrated Agro-environmental Pollution Control and Management, Institute of Eco-environmental and Soil Sciences, Guangdong Academy of Sciences, Guangzhou 510650, China.

出版信息

J Agric Food Chem. 2024 Sep 25;72(38):20862-20871. doi: 10.1021/acs.jafc.4c05055. Epub 2024 Sep 13.

Abstract

The role of insect UDP-glycosyltransferases (UGTs) in the detoxification of insecticides has rarely been reported. A UGT gene was previously found overexpressed in a fenvalerate-resistant strain of . Herein, was cloned, and its involvement in insecticide detoxification was investigated. was highly expressed in the larvae, mainly in the fat body and midgut. Treatment with UGT inhibitors 5-nitrouracil and sulfinpyrazone significantly enhanced the fenvalerate toxicity. Knocking down by RNAi significantly increased the larvae mortality by 17.89%. was further knocked out by CRISPR/Cas9, and a homozygous strain (HD-d) with a C-base deletion at exon 2 was obtained. The sensitivity of HD-d to fenvalerate, deltamethrin, cyantraniliprole, acetamiprid, and lufenuron increased significantly, with sensitivity index increased 2.523-, 2.544-, 2.250-, 2.473-, and 3.556-fold, respectively. These results suggested that was involved in the detoxification of to insecticides mentioned above, shedding light upon further understanding of the detoxification mechanisms of insecticides by insect UGTs.

摘要

昆虫 UDP-糖基转移酶 (UGTs) 在杀虫剂解毒中的作用很少被报道。先前在氰戊菊酯抗性品系 中发现了一个过度表达的 UGT 基因。在此,克隆了 ,并研究了其在杀虫剂解毒中的作用。 在幼虫中高度表达,主要在脂肪体和中肠中表达。用 UGT 抑制剂 5-硝基尿嘧啶和磺吡唑酮处理可显著增强氰戊菊酯的毒性。通过 RNAi 敲低 可使幼虫死亡率显著增加 17.89%。 进一步通过 CRISPR/Cas9 敲除,获得了一个在第 2 外显子处有 C 碱基缺失的纯合株系 (HD-d)。HD-d 对氰戊菊酯、溴氰虫酰胺、虫螨腈、噻虫胺和灭蝇脲的敏感性显著增加,敏感性指数分别增加了 2.523 倍、2.544 倍、2.250 倍、2.473 倍和 3.556 倍。这些结果表明 参与了上述杀虫剂对 的解毒,为进一步了解昆虫 UGTs 对杀虫剂的解毒机制提供了依据。

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